Anti-GFAP Antibody (monoclonal, 3F2)

• Catalog Number: orb623781
• Category: Antibodies
• Description: Anti-GFAP Antibody (monoclonal, 3F2). Tested in IF, IHC, WB applications. This antibody reacts with Human, Mouse, Rat.
• Clonality: Monoclonal
• Species/Host: Mouse
• Isotype: Mouse IgG1
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, Rat
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Immunogen: E.coli-derived human GFAP recombinant protein (Position: Q93-M432). Human GFAP shares 94% amino acid (aa) sequence identity with both mouse and rat GFAP.
• UniProt ID: P14136
• MW: 50 kDa
• Tested applications: IF, IHC, WB
• Application notes: Western blot, 0.1-0.5μg/ml, Mouse, rat Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Rat Immunofluorescence, 5 μg/ml, Rat. Add 0.2ml of distilled water will yield a concentration of 500μg/ml
• Antibody Type: Primary Antibody
• Clone Number: 3F2
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: Cyclin-dependent kinase 1; CDK1; Cell division con
• Note: For research use only

IF analysis of Histone H3 and GFAP using anti-Histone H3 antibody and anti-GFAP antibody. Histone H3 and GFAP was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 µg/mL rabbit anti-Histone H3 antibody and mouse anti-GFAP antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG, Cy3 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IHC analysis of GFAP using anti-GFAP antibody. GFAP was detected in paraffin-embedded section of human glioma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-GFAP Antibody overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of GFAP using anti-GFAP antibody. GFAP was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-GFAP Antibody overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Western blot analysis of GFAP using anti-GFAP antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 ug of sample under reducing conditions. Lane 1: rat brain tissue lysates, Lane 2: mouse brain tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-GFAP antigen affinity purified monoclonal antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for GFAP at approximately 50 KD. The expected band size for GFAP is at 50 KD.